Main Figures and Results

Experimental Design Schematic

Fig.1 Experimental design showing treatments and husbandry events during the course of the study. Symbols indicate when an experimental event occurred at each time point. (1) 260 fish were assigned and acclimated to one of three water temperature groups (e.g., 28°C, 32°C, or 35°C) and reared from 0 to 164 days post fertilization (dpf). (2a) At 164 dpf (or 0 days post exposure; DPE), fecal collections were collected from a random selection of five fish per tank (n = 60). Additionally, histological and wet mount assessments were conducted on selected fish to assess presence of infection and infection burden. (1b) Afterwards, a cohort of fish from each water temperature group were exposed to Pseudocapillaria tomentosa. (3-6) Subsequent fecal samples were collected and histopathological assessments were conducted at 14 dpe (n = 54), (3) 21 dpe (n = 48), (4) 28 dpe (n = 47), and (5) 42 dpe (n = 51).

Results

2) Water temperature shapes gut microbiome structure

Fig. 2 Effects of water temperature on gut microbiomes of parasite unexposed zebrafish. (A) Simpson’s Index of diversity shows that gut microbiome diversity significantly differs between fish reared at 28°C and 35°C water temperatures. (B) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition. The analysis shows that gut microbiome composition significantly differs between fish reared at different water temperatures. (C) Simpson’s Index of diversity shows microbial gut diversity increases with time from 0 days post exposure (dpe) to 42 dpe, irrespective of water temperature. (D) Capscale ordination of gut microbiome composition based on the Bray-Curtis dissimilarity constrained on the main effects of water temperature and time, and their interaction. The analysis shows that shows that gut microbiome composition differs between fish across time depending on water temperature. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate direction of greatest change in the indicated covariates.

2A)

Tables

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GLM

GLM Results
glm(Alpha.Score ~ Temperature); Unexposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−0.754	0.091	−8.309	<0.001	****
Temperature32	0.051	0.127	0.400	≥0.25	ns
Temperature35	0.054	0.129	0.421	≥0.25	ns
Simpson
(Intercept)	−0.101	0.092	−1.103	≥0.25	ns
Temperature32	0.132	0.128	1.030	≥0.25	ns
Temperature35	0.340	0.131	2.591	0.010	*
Richness
(Intercept)	−0.231	0.075	−3.079	0.002	**
Temperature32	0.047	0.105	0.446	≥0.25	ns
Temperature35	−0.473	0.110	−4.299	<0.001	****

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature), type = 2); Unexposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	0.225	2.000	≥0.25	ns
Simpson
Temperature	6.830	2.000	0.033	*
Richness
Temperature	27.512	2.000	<0.001	****

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature); Unexposed fish
term	.y.	group1	group2	estimate	std.error	statistic	adj.p.value	Variable
Shannon
Temperature	Alpha.Score	32	28	0.051	0.127	0.400	≥0.25	Temperature
Temperature	Alpha.Score	35	28	0.054	0.129	0.421	≥0.25	Temperature
Temperature	Alpha.Score	35	32	0.004	0.127	0.029	≥0.25	Temperature
Simpson
Temperature	Alpha.Score	32	28	0.132	0.128	1.030	≥0.25	Temperature
Temperature	Alpha.Score	35	28	0.340	0.131	2.591	0.026	Temperature
Temperature	Alpha.Score	35	32	0.207	0.130	1.596	0.247	Temperature
Richness
Temperature	Alpha.Score	32	28	0.047	0.105	0.446	≥0.25	Temperature
Temperature	Alpha.Score	35	28	−0.473	0.110	−4.299	<0.001	Temperature
Temperature	Alpha.Score	35	32	−0.520	0.109	−4.768	<0.001	Temperature

2B)

Tables

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Capscale

ADONIS2
adonis2(Beta Distance ~ Temperature); Unexposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	2.294	0.133	12.900	0.001	***
Residual	168.000	14.936	0.867	NA	NA	NA
Total	170.000	17.229	1.000	NA	NA	NA
canberra
Temperature	2.000	3.862	0.099	9.178	0.001	***
Residual	168.000	35.344	0.901	NA	NA	NA
Total	170.000	39.206	1.000	NA	NA	NA
gunifrac
Temperature	2.000	1.422	0.102	9.566	0.001	***
Residual	168.000	12.485	0.898	NA	NA	NA
Total	170.000	13.907	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Unexposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temperature	2.000	0.008	0.004	0.386	≥0.25	ns
Residual	168.000	1.771	0.011	NA	NA	NA
canberra
Temperature	2.000	0.008	0.004	0.873	≥0.25	ns
Residual	168.000	0.759	0.005	NA	NA	NA
gunifrac
Temperature	2.000	0.005	0.003	0.369	≥0.25	ns
Residual	168.000	1.143	0.007	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature); Unexposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temperature	32	28	−0.011	−0.056	0.034	≥0.25	ns
Distance	Temperature	35	28	0.005	−0.041	0.051	≥0.25	ns
Distance	Temperature	35	32	0.016	−0.029	0.062	≥0.25	ns
canberra
Distance	Temperature	32	28	−0.001	−0.031	0.028	≥0.25	ns
Distance	Temperature	35	28	0.014	−0.016	0.044	≥0.25	ns
Distance	Temperature	35	32	0.015	−0.015	0.045	≥0.25	ns
gunifrac
Distance	Temperature	32	28	−0.013	−0.049	0.023	≥0.25	ns
Distance	Temperature	35	28	−0.004	−0.040	0.033	≥0.25	ns
Distance	Temperature	35	32	0.009	−0.027	0.046	≥0.25	ns

2C)

Tables

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GLM

GLM Results
glm(Alpha.Score ~ Temperature*DPE); Unexposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−1.074	0.138	−7.754	<0.001	****
Temperature32	0.075	0.193	0.386	≥0.25	ns
Temperature35	0.154	0.193	0.797	≥0.25	ns
DPE	0.018	0.006	3.086	0.002	**
Temperature32:DPE	−0.001	0.008	−0.170	≥0.25	ns
Temperature35:DPE	−0.005	0.008	−0.585	≥0.25	ns
Simpson
(Intercept)	−0.433	0.134	−3.245	0.001	**
Temperature32	0.157	0.186	0.841	≥0.25	ns
Temperature35	0.408	0.187	2.177	0.031	*
DPE	0.019	0.006	3.293	0.001	**
Temperature32:DPE	−0.001	0.008	−0.173	≥0.25	ns
Temperature35:DPE	−0.003	0.008	−0.343	≥0.25	ns
Richness
(Intercept)	−0.651	0.108	−6.017	<0.001	****
Temperature32	0.189	0.150	1.257	0.211	ns
Temperature35	−0.059	0.154	−0.383	≥0.25	ns
DPE	0.024	0.005	5.137	<0.001	****
Temperature32:DPE	−0.008	0.006	−1.251	0.213	ns
Temperature35:DPE	−0.024	0.007	−3.476	<0.001	***

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Unexposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	0.322	2.000	≥0.25	ns
DPE	22.895	1.000	<0.001	****
Temperature:DPE	0.363	2.000	≥0.25	ns
Simpson
Temperature	8.551	2.000	0.014	*
DPE	28.161	1.000	<0.001	****
Temperature:DPE	0.118	2.000	≥0.25	ns
Richness
Temperature	31.759	2.000	<0.001	****
DPE	27.201	1.000	<0.001	****
Temperature:DPE	12.460	2.000	0.002	**

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Unexposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	DPE	14	0	1.065	0.294	3.625	0.003	DPE	28
28	Alpha.Score	DPE	21	0	0.722	0.281	2.566	0.076	DPE	28
28	Alpha.Score	DPE	28	0	1.187	0.274	4.335	<0.001	DPE	28
28	Alpha.Score	DPE	42	0	0.548	0.296	1.853	≥0.25	DPE	28
28	Alpha.Score	DPE	21	14	−0.343	0.317	−1.083	≥0.25	DPE	28
28	Alpha.Score	DPE	28	14	0.122	0.310	0.392	≥0.25	DPE	28
28	Alpha.Score	DPE	42	14	−0.517	0.330	−1.567	≥0.25	DPE	28
28	Alpha.Score	DPE	28	21	0.465	0.299	1.558	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	−0.174	0.319	−0.545	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	−0.639	0.312	−2.045	0.243	DPE	28
32	Alpha.Score	DPE	14	0	0.006	0.250	0.022	≥0.25	DPE	32
32	Alpha.Score	DPE	21	0	0.112	0.247	0.452	≥0.25	DPE	32
32	Alpha.Score	DPE	28	0	0.420	0.248	1.695	≥0.25	DPE	32
32	Alpha.Score	DPE	42	0	0.694	0.236	2.943	0.027	DPE	32
32	Alpha.Score	DPE	21	14	0.106	0.286	0.371	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	0.414	0.287	1.445	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	0.688	0.276	2.490	0.092	DPE	32
32	Alpha.Score	DPE	28	21	0.308	0.283	1.087	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.582	0.273	2.131	0.205	DPE	32
32	Alpha.Score	DPE	42	28	0.274	0.274	1.001	≥0.25	DPE	32
35	Alpha.Score	DPE	14	0	−0.006	0.210	−0.027	≥0.25	DPE	35
35	Alpha.Score	DPE	21	0	−0.249	0.218	−1.145	≥0.25	DPE	35
35	Alpha.Score	DPE	28	0	0.232	0.204	1.138	≥0.25	DPE	35
35	Alpha.Score	DPE	42	0	0.661	0.207	3.194	0.012	DPE	35
35	Alpha.Score	DPE	21	14	−0.243	0.253	−0.962	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	0.238	0.242	0.985	≥0.25	DPE	35
35	Alpha.Score	DPE	42	14	0.667	0.244	2.733	0.049	DPE	35
35	Alpha.Score	DPE	28	21	0.481	0.248	1.938	≥0.25	DPE	35
35	Alpha.Score	DPE	42	21	0.910	0.251	3.628	0.003	DPE	35
35	Alpha.Score	DPE	42	28	0.429	0.239	1.792	≥0.25	DPE	35
Simpson
28	Alpha.Score	DPE	14	0	1.256	0.273	4.611	<0.001	DPE	28
28	Alpha.Score	DPE	21	0	0.761	0.247	3.076	0.018	DPE	28
28	Alpha.Score	DPE	28	0	1.334	0.254	5.241	<0.001	DPE	28
28	Alpha.Score	DPE	42	0	0.514	0.257	2.002	≥0.25	DPE	28
28	Alpha.Score	DPE	21	14	−0.496	0.302	−1.644	≥0.25	DPE	28
28	Alpha.Score	DPE	28	14	0.077	0.308	0.251	≥0.25	DPE	28
28	Alpha.Score	DPE	42	14	−0.742	0.310	−2.398	0.114	DPE	28
28	Alpha.Score	DPE	28	21	0.573	0.285	2.008	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	−0.246	0.288	−0.857	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	−0.819	0.294	−2.789	0.042	DPE	28
32	Alpha.Score	DPE	14	0	0.065	0.249	0.261	≥0.25	DPE	32
32	Alpha.Score	DPE	21	0	0.142	0.249	0.569	≥0.25	DPE	32
32	Alpha.Score	DPE	28	0	0.592	0.261	2.272	0.152	DPE	32
32	Alpha.Score	DPE	42	0	0.724	0.254	2.850	0.035	DPE	32
32	Alpha.Score	DPE	21	14	0.077	0.287	0.267	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	0.527	0.297	1.773	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	0.659	0.292	2.260	0.156	DPE	32
32	Alpha.Score	DPE	28	21	0.451	0.297	1.517	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.582	0.291	1.999	≥0.25	DPE	32
32	Alpha.Score	DPE	42	28	0.132	0.302	0.436	≥0.25	DPE	32
35	Alpha.Score	DPE	14	0	0.149	0.219	0.683	≥0.25	DPE	35
35	Alpha.Score	DPE	21	0	−0.167	0.218	−0.766	≥0.25	DPE	35
35	Alpha.Score	DPE	28	0	0.413	0.222	1.858	≥0.25	DPE	35
35	Alpha.Score	DPE	42	0	0.836	0.244	3.431	0.005	DPE	35
35	Alpha.Score	DPE	21	14	−0.316	0.257	−1.232	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	0.263	0.260	1.013	≥0.25	DPE	35
35	Alpha.Score	DPE	42	14	0.687	0.279	2.465	0.097	DPE	35
35	Alpha.Score	DPE	28	21	0.579	0.260	2.233	0.165	DPE	35
35	Alpha.Score	DPE	42	21	1.003	0.278	3.604	0.003	DPE	35
35	Alpha.Score	DPE	42	28	0.424	0.282	1.506	≥0.25	DPE	35
Richness
28	Alpha.Score	DPE	14	0	0.432	0.236	1.836	≥0.25	DPE	28
28	Alpha.Score	DPE	21	0	0.550	0.218	2.526	0.084	DPE	28
28	Alpha.Score	DPE	28	0	0.767	0.218	3.525	0.004	DPE	28
28	Alpha.Score	DPE	42	0	0.970	0.227	4.278	<0.001	DPE	28
28	Alpha.Score	DPE	21	14	0.118	0.263	0.449	≥0.25	DPE	28
28	Alpha.Score	DPE	28	14	0.335	0.262	1.275	≥0.25	DPE	28
28	Alpha.Score	DPE	42	14	0.537	0.270	1.990	≥0.25	DPE	28
28	Alpha.Score	DPE	28	21	0.217	0.247	0.880	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	0.419	0.255	1.647	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	0.203	0.254	0.796	≥0.25	DPE	28
32	Alpha.Score	DPE	14	0	−0.170	0.177	−0.965	≥0.25	DPE	32
32	Alpha.Score	DPE	21	0	−0.026	0.175	−0.147	≥0.25	DPE	32
32	Alpha.Score	DPE	28	0	0.187	0.180	1.041	≥0.25	DPE	32
32	Alpha.Score	DPE	42	0	0.769	0.176	4.359	<0.001	DPE	32
32	Alpha.Score	DPE	21	14	0.145	0.204	0.711	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	0.358	0.208	1.721	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	0.939	0.205	4.585	<0.001	DPE	32
32	Alpha.Score	DPE	28	21	0.213	0.206	1.031	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.794	0.203	3.906	<0.001	DPE	32
32	Alpha.Score	DPE	42	28	0.581	0.208	2.801	0.040	DPE	32
35	Alpha.Score	DPE	14	0	−0.560	0.214	−2.612	0.067	DPE	35
35	Alpha.Score	DPE	21	0	−0.551	0.214	−2.578	0.073	DPE	35
35	Alpha.Score	DPE	28	0	0.110	0.196	0.561	≥0.25	DPE	35
35	Alpha.Score	DPE	42	0	0.007	0.206	0.036	≥0.25	DPE	35
35	Alpha.Score	DPE	21	14	0.008	0.259	0.032	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	0.670	0.245	2.733	0.048	DPE	35
35	Alpha.Score	DPE	42	14	0.567	0.253	2.240	0.162	DPE	35
35	Alpha.Score	DPE	28	21	0.661	0.245	2.702	0.053	DPE	35
35	Alpha.Score	DPE	42	21	0.559	0.253	2.210	0.173	DPE	35
35	Alpha.Score	DPE	42	28	−0.103	0.238	−0.431	≥0.25	DPE	35

2D)

Tables

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Capscale

ADONIS2
adonis2(Beta Distance ~ Temperature*DPE); Unexposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	2.294	0.133	16.443	0.001	***
DPE	1.000	2.531	0.147	36.284	0.001	***
Temperature:DPE	2.000	0.896	0.052	6.425	0.001	***
Residual	165.000	11.509	0.668	NA	NA	NA
Total	170.000	17.229	1.000	NA	NA	NA
canberra
Temperature	2.000	3.862	0.099	10.667	0.001	***
DPE	1.000	3.655	0.093	20.192	0.001	***
Temperature:DPE	2.000	1.820	0.046	5.026	0.001	***
Residual	165.000	29.869	0.762	NA	NA	NA
Total	170.000	39.206	1.000	NA	NA	NA
gunifrac
Temperature	2.000	1.422	0.102	11.984	0.001	***
DPE	1.000	2.123	0.153	35.785	0.001	***
Temperature:DPE	2.000	0.574	0.041	4.840	0.001	***
Residual	165.000	9.788	0.704	NA	NA	NA
Total	170.000	13.907	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Unexposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temp.DPE	14.000	0.290	0.021	2.348	0.006	**
Residual	156.000	1.376	0.009	NA	NA	NA
canberra
Temp.DPE	14.000	0.250	0.018	3.344	<0.001	***
Residual	156.000	0.831	0.005	NA	NA	NA
gunifrac
Temp.DPE	14.000	0.249	0.018	2.986	<0.001	***
Residual	156.000	0.929	0.006	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*DPE); Unexposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temp.DPE	28°C__14DPE	28°C__0DPE	0.024	−0.112	0.159	≥0.25	ns
Distance	Temp.DPE	28°C__21DPE	28°C__0DPE	0.052	−0.074	0.177	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__0DPE	0.007	−0.118	0.133	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__0DPE	−0.017	−0.147	0.113	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__0DPE	0.097	−0.005	0.199	0.085	ns
Distance	Temp.DPE	32°C__14DPE	28°C__0DPE	0.041	−0.085	0.166	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__0DPE	−0.018	−0.143	0.108	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__0DPE	0.028	−0.102	0.158	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__0DPE	0.017	−0.108	0.142	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__0DPE	0.044	−0.058	0.146	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__0DPE	0.004	−0.126	0.134	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__0DPE	−0.004	−0.134	0.126	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__0DPE	0.033	−0.097	0.162	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__0DPE	0.146	0.011	0.282	0.021	*
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	0.028	−0.126	0.181	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	−0.016	−0.170	0.137	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	−0.040	−0.198	0.117	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__14DPE	0.073	−0.062	0.209	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	0.017	−0.136	0.171	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.041	−0.195	0.112	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	0.005	−0.153	0.162	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.006	−0.160	0.147	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__14DPE	0.020	−0.115	0.156	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.019	−0.176	0.138	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.028	−0.185	0.129	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	0.009	−0.148	0.166	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	0.123	−0.039	0.284	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	−0.044	−0.189	0.101	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	−0.068	−0.217	0.081	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__21DPE	0.045	−0.080	0.171	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	−0.011	−0.156	0.134	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	−0.069	−0.214	0.076	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	−0.023	−0.172	0.125	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	−0.034	−0.179	0.110	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__21DPE	−0.008	−0.133	0.118	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	−0.047	−0.196	0.102	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	−0.056	−0.205	0.093	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.019	−0.168	0.130	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	0.095	−0.059	0.248	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	−0.024	−0.173	0.125	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__28DPE	0.089	−0.036	0.215	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	0.033	−0.111	0.178	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	−0.025	−0.170	0.120	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	0.021	−0.128	0.169	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	0.010	−0.135	0.154	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__28DPE	0.036	−0.089	0.162	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	−0.003	−0.152	0.146	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	−0.012	−0.161	0.137	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	0.025	−0.124	0.174	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	0.139	−0.015	0.292	0.125	ns
Distance	Temp.DPE	32°C__0DPE	28°C__42DPE	0.114	−0.016	0.243	0.161	ns
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	0.057	−0.091	0.206	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	−0.001	−0.150	0.148	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	0.045	−0.108	0.197	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	0.034	−0.115	0.183	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__42DPE	0.061	−0.069	0.191	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	0.021	−0.131	0.174	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	0.012	−0.140	0.165	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	0.049	−0.103	0.202	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	0.163	0.006	0.320	0.034	*
Distance	Temp.DPE	32°C__14DPE	32°C__0DPE	−0.056	−0.182	0.069	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__0DPE	−0.114	−0.240	0.011	0.115	ns
Distance	Temp.DPE	32°C__28DPE	32°C__0DPE	−0.069	−0.199	0.061	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__0DPE	−0.080	−0.205	0.046	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__0DPE	−0.053	−0.155	0.049	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__0DPE	−0.092	−0.222	0.037	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__0DPE	−0.101	−0.231	0.029	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__0DPE	−0.064	−0.194	0.066	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__0DPE	0.049	−0.086	0.185	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.058	−0.203	0.086	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.013	−0.161	0.136	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.024	−0.168	0.121	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__14DPE	0.003	−0.122	0.129	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.036	−0.185	0.112	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.045	−0.194	0.104	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.008	−0.157	0.141	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	0.105	−0.048	0.259	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	0.046	−0.103	0.194	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	0.035	−0.110	0.179	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__21DPE	0.061	−0.064	0.187	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	0.022	−0.127	0.171	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	0.013	−0.136	0.162	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	0.050	−0.099	0.199	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	0.164	0.010	0.317	0.025	*
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	−0.011	−0.160	0.138	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__28DPE	0.016	−0.114	0.146	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	−0.024	−0.176	0.129	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	−0.033	−0.185	0.120	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	0.004	−0.148	0.157	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	0.118	−0.039	0.275	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__42DPE	0.027	−0.099	0.152	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	−0.013	−0.161	0.136	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	−0.021	−0.170	0.127	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	0.015	−0.133	0.164	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	0.129	−0.024	0.283	0.209	ns
Distance	Temp.DPE	35°C__14DPE	35°C__0DPE	−0.039	−0.169	0.090	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__0DPE	−0.048	−0.178	0.082	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__0DPE	−0.011	−0.141	0.119	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__0DPE	0.102	−0.033	0.238	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	−0.009	−0.161	0.144	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	0.028	−0.124	0.181	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	0.142	−0.016	0.299	0.127	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	0.037	−0.116	0.190	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	0.151	−0.007	0.308	0.076	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.114	−0.044	0.271	≥0.25	ns
canberra
Distance	Temp.DPE	28°C__14DPE	28°C__0DPE	−0.033	−0.138	0.072	≥0.25	ns
Distance	Temp.DPE	28°C__21DPE	28°C__0DPE	−0.005	−0.103	0.092	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__0DPE	−0.074	−0.171	0.024	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__0DPE	−0.069	−0.170	0.032	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__0DPE	0.035	−0.045	0.115	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__0DPE	−0.021	−0.119	0.076	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__0DPE	−0.070	−0.167	0.028	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__0DPE	−0.056	−0.157	0.045	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__0DPE	−0.068	−0.165	0.030	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__0DPE	0.008	−0.071	0.088	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__0DPE	−0.041	−0.142	0.060	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__0DPE	−0.047	−0.148	0.054	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__0DPE	−0.052	−0.153	0.049	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__0DPE	0.045	−0.061	0.150	≥0.25	ns
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	0.028	−0.091	0.147	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	−0.040	−0.160	0.079	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	−0.035	−0.158	0.087	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__14DPE	0.068	−0.037	0.174	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	0.012	−0.107	0.131	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.037	−0.156	0.083	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	−0.023	−0.145	0.099	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.034	−0.154	0.085	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__14DPE	0.042	−0.064	0.147	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.008	−0.130	0.115	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.013	−0.136	0.109	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	−0.019	−0.141	0.103	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	0.078	−0.048	0.204	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	−0.068	−0.181	0.044	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	−0.063	−0.179	0.052	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__21DPE	0.040	−0.057	0.138	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	−0.016	−0.128	0.097	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	−0.065	−0.177	0.048	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	−0.051	−0.167	0.065	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	−0.062	−0.175	0.050	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__21DPE	0.014	−0.084	0.111	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	−0.036	−0.151	0.080	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	−0.041	−0.157	0.074	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.047	−0.162	0.069	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	0.050	−0.070	0.169	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	0.005	−0.110	0.121	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__28DPE	0.109	0.011	0.206	0.014	*
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	0.052	−0.060	0.165	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	0.004	−0.109	0.116	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	0.017	−0.098	0.133	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	0.006	−0.106	0.119	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__28DPE	0.082	−0.015	0.180	0.206	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	0.033	−0.083	0.148	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	0.027	−0.088	0.143	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	0.021	−0.094	0.137	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	0.118	−0.001	0.237	0.055	ns
Distance	Temp.DPE	32°C__0DPE	28°C__42DPE	0.104	0.003	0.205	0.038	*
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	0.047	−0.068	0.163	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	−0.001	−0.117	0.114	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	0.012	−0.106	0.131	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	0.001	−0.115	0.117	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__42DPE	0.077	−0.024	0.178	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	0.028	−0.091	0.146	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	0.022	−0.097	0.141	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	0.016	−0.102	0.135	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	0.113	−0.009	0.235	0.104	ns
Distance	Temp.DPE	32°C__14DPE	32°C__0DPE	−0.056	−0.154	0.041	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__0DPE	−0.105	−0.202	−0.007	0.022	*
Distance	Temp.DPE	32°C__28DPE	32°C__0DPE	−0.091	−0.192	0.010	0.125	ns
Distance	Temp.DPE	32°C__42DPE	32°C__0DPE	−0.103	−0.200	−0.005	0.029	*
Distance	Temp.DPE	35°C__0DPE	32°C__0DPE	−0.027	−0.106	0.053	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__0DPE	−0.076	−0.177	0.025	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__0DPE	−0.082	−0.183	0.019	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__0DPE	−0.087	−0.188	0.014	0.175	ns
Distance	Temp.DPE	35°C__42DPE	32°C__0DPE	0.009	−0.096	0.115	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.049	−0.161	0.064	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.035	−0.151	0.081	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.046	−0.159	0.066	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__14DPE	0.030	−0.068	0.127	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.020	−0.135	0.096	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.025	−0.141	0.090	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.031	−0.147	0.085	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	0.066	−0.054	0.185	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	0.014	−0.102	0.129	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	0.002	−0.110	0.115	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__21DPE	0.078	−0.019	0.176	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	0.029	−0.087	0.145	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	0.023	−0.092	0.139	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	0.018	−0.098	0.133	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	0.114	−0.005	0.234	0.076	ns
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	−0.011	−0.127	0.104	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__28DPE	0.065	−0.036	0.166	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	0.015	−0.103	0.134	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	0.010	−0.109	0.128	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	0.004	−0.115	0.123	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	0.101	−0.022	0.223	0.237	ns
Distance	Temp.DPE	35°C__0DPE	32°C__42DPE	0.076	−0.021	0.173	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	0.027	−0.089	0.142	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	0.021	−0.095	0.137	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	0.015	−0.100	0.131	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	0.112	−0.007	0.231	0.091	ns
Distance	Temp.DPE	35°C__14DPE	35°C__0DPE	−0.049	−0.150	0.052	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__0DPE	−0.055	−0.156	0.046	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__0DPE	−0.061	−0.162	0.040	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__0DPE	0.036	−0.069	0.141	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	−0.006	−0.124	0.113	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	−0.011	−0.130	0.107	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	0.085	−0.037	0.208	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	−0.006	−0.124	0.113	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	0.091	−0.031	0.213	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.097	−0.026	0.219	≥0.25	ns
gunifrac
Distance	Temp.DPE	28°C__14DPE	28°C__0DPE	−0.005	−0.116	0.107	≥0.25	ns
Distance	Temp.DPE	28°C__21DPE	28°C__0DPE	0.038	−0.065	0.141	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__0DPE	−0.010	−0.113	0.093	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__0DPE	−0.033	−0.140	0.073	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__0DPE	0.067	−0.017	0.151	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__0DPE	0.018	−0.085	0.121	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__0DPE	−0.023	−0.126	0.080	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__0DPE	−0.014	−0.121	0.093	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__0DPE	−0.026	−0.129	0.077	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__0DPE	0.029	−0.055	0.113	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__0DPE	−0.015	−0.122	0.092	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__0DPE	−0.020	−0.127	0.087	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__0DPE	−0.065	−0.171	0.042	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__0DPE	0.091	−0.020	0.202	0.245	ns
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	0.042	−0.084	0.168	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	−0.005	−0.131	0.121	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	−0.029	−0.158	0.100	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__14DPE	0.072	−0.040	0.183	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	0.022	−0.104	0.148	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.019	−0.145	0.107	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	−0.009	−0.138	0.120	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.021	−0.147	0.105	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__14DPE	0.033	−0.078	0.145	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.010	−0.139	0.119	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.015	−0.145	0.114	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	−0.060	−0.189	0.069	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	0.096	−0.037	0.229	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	−0.047	−0.166	0.072	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	−0.071	−0.193	0.051	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__21DPE	0.029	−0.074	0.132	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	−0.020	−0.139	0.099	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	−0.061	−0.180	0.058	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	−0.051	−0.174	0.071	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	−0.063	−0.182	0.056	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__21DPE	−0.009	−0.112	0.094	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	−0.052	−0.175	0.070	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	−0.058	−0.180	0.065	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.102	−0.224	0.020	0.216	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	0.054	−0.073	0.180	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	−0.024	−0.146	0.098	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__28DPE	0.077	−0.026	0.180	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	0.027	−0.092	0.146	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	−0.014	−0.133	0.105	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	−0.004	−0.126	0.118	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	−0.016	−0.135	0.103	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__28DPE	0.038	−0.065	0.142	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	−0.005	−0.127	0.117	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	−0.010	−0.132	0.112	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	−0.055	−0.177	0.067	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	0.101	−0.025	0.227	≥0.25	ns
Distance	Temp.DPE	32°C__0DPE	28°C__42DPE	0.100	−0.006	0.207	0.089	ns
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	0.051	−0.071	0.173	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	0.010	−0.112	0.132	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	0.020	−0.106	0.145	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	0.008	−0.114	0.130	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	28°C__42DPE	0.062	−0.045	0.169	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	0.019	−0.107	0.144	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	0.014	−0.112	0.139	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	−0.031	−0.157	0.094	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	0.125	−0.005	0.254	0.071	ns
Distance	Temp.DPE	32°C__14DPE	32°C__0DPE	−0.049	−0.152	0.054	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__0DPE	−0.090	−0.193	0.013	0.155	ns
Distance	Temp.DPE	32°C__28DPE	32°C__0DPE	−0.081	−0.188	0.026	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__0DPE	−0.093	−0.196	0.010	0.130	ns
Distance	Temp.DPE	35°C__0DPE	32°C__0DPE	−0.038	−0.122	0.046	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__0DPE	−0.082	−0.189	0.025	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__0DPE	−0.087	−0.194	0.020	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__0DPE	−0.132	−0.238	−0.025	0.003	**
Distance	Temp.DPE	35°C__42DPE	32°C__0DPE	0.024	−0.087	0.135	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.041	−0.160	0.078	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.031	−0.154	0.091	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.043	−0.162	0.076	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__14DPE	0.011	−0.092	0.114	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.032	−0.155	0.090	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.037	−0.160	0.085	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.082	−0.204	0.040	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	0.074	−0.053	0.200	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	0.010	−0.113	0.132	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	−0.002	−0.121	0.117	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__21DPE	0.052	−0.051	0.155	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	0.009	−0.114	0.131	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	0.004	−0.119	0.126	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	−0.041	−0.163	0.081	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	0.115	−0.011	0.241	0.119	ns
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	−0.012	−0.134	0.110	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__28DPE	0.043	−0.064	0.149	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	−0.001	−0.126	0.124	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	−0.006	−0.132	0.119	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	−0.051	−0.176	0.075	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	0.105	−0.024	0.234	≥0.25	ns
Distance	Temp.DPE	35°C__0DPE	32°C__42DPE	0.054	−0.049	0.157	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	0.011	−0.111	0.133	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	0.006	−0.117	0.128	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	−0.039	−0.161	0.083	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	0.117	−0.009	0.243	0.102	ns
Distance	Temp.DPE	35°C__14DPE	35°C__0DPE	−0.044	−0.150	0.063	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__0DPE	−0.049	−0.155	0.058	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__0DPE	−0.093	−0.200	0.013	0.160	ns
Distance	Temp.DPE	35°C__42DPE	35°C__0DPE	0.062	−0.049	0.174	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	−0.005	−0.130	0.120	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	−0.050	−0.175	0.076	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	0.106	−0.023	0.235	0.244	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	−0.045	−0.170	0.081	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	0.111	−0.018	0.240	0.180	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.156	0.027	0.285	0.005	**

3) Parasitic worm burden differs by water temperature

Fig. 3 Histological sections stained with ### stain in zebrafish exposed to Pseudocapillaria tomentosa examined at ### days post exposure. (A) Nematodes in intestinal lumen. Arrow = ###. (B) Infection outcome analysis of fish exposed to P. tomentosa (n = 89) by temperature. Fish reared at 28°C and 32°C water temperatures had significantly different infection burden to fish reared at 35°C water temperature. Only one fish reared at 35°C was identified as being positively infected by wet mount.

*3A)

3B)

4) Water temperature differentially stratifies gut microbiome communities of parasite exposed fish

Fig. 4 Effects of parasite exposure and water temperature on gut microbiomes of zebrafish. (A) Simpson’s Index of diversity shows that gut microbiome diversity significantly differs between fish reared at 28°C water temperature to fish reared at 32°C and 35°C water temperatures. (B) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition. The analysis shows that gut microbiome composition significantly differs between parasite exposed fish reared at different water temperatures. (C) Simpson’s Index of diversity shows microbial gut diversity decreases with time from 0 days post exposure (dpe) to 42 dpe in parasite exposed fish reared at 28°C water temperature. (D) Capscale ordination of gut microbiome composition based on the Canberra dissimilarity constrained on the main effects of water temperature and time, and their interaction. The analysis shows that shows that gut microbiome composition differs between parasite exposed fish across time depending on water temperature. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate direction of greatest change in the indicated covariates.

4A)

Tables

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GLM

GLM Results
glm(Alpha.Score ~ Temperature); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−0.936	0.233	−4.021	<0.001	***
Temperature32	0.909	0.301	3.017	0.003	**
Temperature35	0.516	0.296	1.741	0.085	ns
Simpson
(Intercept)	−1.002	0.243	−4.130	<0.001	****
Temperature32	1.057	0.312	3.384	0.001	**
Temperature35	0.772	0.306	2.525	0.013	*
Richness
(Intercept)	−0.681	0.190	−3.583	<0.001	***
Temperature32	0.703	0.251	2.800	0.006	**
Temperature35	0.242	0.247	0.978	≥0.25	ns

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature), type = 2); Exposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	9.440	2.000	0.009	**
Simpson
Temperature	12.497	2.000	0.002	**
Richness
Temperature	8.575	2.000	0.014	*

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature); Exposed fish
term	.y.	group1	group2	estimate	std.error	statistic	adj.p.value	Variable
Shannon
Temperature	Alpha.Score	32	28	0.909	0.301	3.017	0.007	Temperature
Temperature	Alpha.Score	35	28	0.516	0.296	1.741	0.190	Temperature
Temperature	Alpha.Score	35	32	−0.393	0.265	−1.482	≥0.25	Temperature
Simpson
Temperature	Alpha.Score	32	28	1.057	0.312	3.384	0.002	Temperature
Temperature	Alpha.Score	35	28	0.772	0.306	2.525	0.031	Temperature
Temperature	Alpha.Score	35	32	−0.285	0.270	−1.054	≥0.25	Temperature
Richness
Temperature	Alpha.Score	32	28	0.703	0.251	2.800	0.014	Temperature
Temperature	Alpha.Score	35	28	0.242	0.247	0.978	≥0.25	Temperature
Temperature	Alpha.Score	35	32	−0.461	0.228	−2.028	0.105	Temperature

4B)

Tables

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Capscale

ADONIS2
adonis2(Beta Distance ~ Temperature); Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	5.218	0.251	14.447	0.001	***
Residual	86.000	15.530	0.749	NA	NA	NA
Total	88.000	20.748	1.000	NA	NA	NA
canberra
Temperature	2.000	3.573	0.117	5.701	0.001	***
Residual	86.000	26.947	0.883	NA	NA	NA
Total	88.000	30.520	1.000	NA	NA	NA
gunifrac
Temperature	2.000	3.363	0.212	11.590	0.001	***
Residual	86.000	12.478	0.788	NA	NA	NA
Total	88.000	15.841	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Exposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temperature	2.000	0.364	0.182	8.708	<0.001	***
Residual	86.000	1.796	0.021	NA	NA	NA
canberra
Temperature	2.000	0.314	0.157	17.419	<0.001	****
Residual	86.000	0.774	0.009	NA	NA	NA
gunifrac
Temperature	2.000	0.310	0.155	13.567	<0.001	****
Residual	86.000	0.983	0.011	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature); Exposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temperature	32	28	−0.043	−0.136	0.051	≥0.25	ns
Distance	Temperature	35	28	−0.150	−0.241	−0.059	<0.001	***
Distance	Temperature	35	32	−0.108	−0.194	−0.021	0.011	*
canberra
Distance	Temperature	32	28	−0.110	−0.171	−0.049	<0.001	***
Distance	Temperature	35	28	−0.144	−0.204	−0.085	<0.001	****
Distance	Temperature	35	32	−0.034	−0.091	0.022	≥0.25	ns
gunifrac
Distance	Temperature	32	28	−0.056	−0.125	0.013	0.133	ns
Distance	Temperature	35	28	−0.143	−0.211	−0.076	<0.001	****
Distance	Temperature	35	32	−0.087	−0.151	−0.023	0.005	**

4C)

Tables

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GLM

GLM Results
glm(Alpha.Score ~ Temperature*DPE); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−0.134	0.594	−0.226	≥0.25	ns
Temperature32	0.228	0.774	0.295	≥0.25	ns
Temperature35	−0.428	0.777	−0.551	≥0.25	ns
DPE	−0.031	0.022	−1.425	0.158	ns
Temperature32:DPE	0.026	0.028	0.917	≥0.25	ns
Temperature35:DPE	0.036	0.028	1.300	0.197	ns
Simpson
(Intercept)	−0.282	0.617	−0.456	≥0.25	ns
Temperature32	0.336	0.801	0.420	≥0.25	ns
Temperature35	−0.172	0.799	−0.215	≥0.25	ns
DPE	−0.028	0.022	−1.234	0.221	ns
Temperature32:DPE	0.028	0.029	0.946	≥0.25	ns
Temperature35:DPE	0.036	0.029	1.261	0.211	ns
Richness
(Intercept)	0.316	0.480	0.657	≥0.25	ns
Temperature32	−0.034	0.636	−0.054	≥0.25	ns
Temperature35	−0.596	0.638	−0.934	≥0.25	ns
DPE	−0.038	0.017	−2.191	0.031	*
Temperature32:DPE	0.028	0.023	1.199	0.234	ns
Temperature35:DPE	0.032	0.023	1.405	0.164	ns

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Exposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	8.789	2.000	0.012	*
DPE	0.523	1.000	≥0.25	ns
Temperature:DPE	1.759	2.000	≥0.25	ns
Simpson
Temperature	11.871	2.000	0.003	**
DPE	0.120	1.000	≥0.25	ns
Temperature:DPE	1.684	2.000	≥0.25	ns
Richness
Temperature	7.911	2.000	0.019	*
DPE	3.435	1.000	0.064	ns
Temperature:DPE	2.255	2.000	≥0.25	ns

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Exposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	DPE	21	14	−3.311	1.822	−1.817	0.246	DPE	28
28	Alpha.Score	DPE	28	14	−2.027	1.023	−1.982	0.178	DPE	28
28	Alpha.Score	DPE	42	14	−1.024	0.705	−1.452	≥0.25	DPE	28
28	Alpha.Score	DPE	28	21	1.284	1.982	0.648	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	2.287	1.838	1.245	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	1.004	1.050	0.956	≥0.25	DPE	28
32	Alpha.Score	DPE	21	14	−0.536	0.537	−0.999	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	0.056	0.558	0.099	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	−0.271	0.532	−0.509	≥0.25	DPE	32
32	Alpha.Score	DPE	28	21	0.592	0.606	0.977	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.266	0.581	0.457	≥0.25	DPE	32
32	Alpha.Score	DPE	42	28	−0.326	0.601	−0.543	≥0.25	DPE	32
35	Alpha.Score	DPE	21	14	−0.246	0.270	−0.909	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	−0.292	0.271	−1.075	≥0.25	DPE	35
35	Alpha.Score	DPE	42	14	0.097	0.258	0.377	≥0.25	DPE	35
35	Alpha.Score	DPE	28	21	−0.046	0.283	−0.162	≥0.25	DPE	35
35	Alpha.Score	DPE	42	21	0.343	0.270	1.273	≥0.25	DPE	35
35	Alpha.Score	DPE	42	28	0.389	0.271	1.438	≥0.25	DPE	35
Simpson
28	Alpha.Score	DPE	21	14	−3.141	1.820	−1.725	≥0.25	DPE	28
28	Alpha.Score	DPE	28	14	−1.994	1.072	−1.861	0.226	DPE	28
28	Alpha.Score	DPE	42	14	−0.925	0.721	−1.283	≥0.25	DPE	28
28	Alpha.Score	DPE	28	21	1.147	2.001	0.573	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	2.215	1.837	1.206	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	1.069	1.100	0.971	≥0.25	DPE	28
32	Alpha.Score	DPE	21	14	−0.416	0.550	−0.756	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	0.275	0.580	0.474	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	−0.138	0.546	−0.252	≥0.25	DPE	32
32	Alpha.Score	DPE	28	21	0.691	0.626	1.103	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.278	0.595	0.467	≥0.25	DPE	32
32	Alpha.Score	DPE	42	28	−0.413	0.623	−0.662	≥0.25	DPE	32
35	Alpha.Score	DPE	21	14	−0.374	0.297	−1.258	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	−0.285	0.296	−0.966	≥0.25	DPE	35
35	Alpha.Score	DPE	42	14	0.142	0.284	0.502	≥0.25	DPE	35
35	Alpha.Score	DPE	28	21	0.088	0.308	0.287	≥0.25	DPE	35
35	Alpha.Score	DPE	42	21	0.516	0.297	1.738	≥0.25	DPE	35
35	Alpha.Score	DPE	42	28	0.428	0.295	1.448	≥0.25	DPE	35
Richness
28	Alpha.Score	DPE	21	14	−2.219	1.016	−2.185	0.123	DPE	28
28	Alpha.Score	DPE	28	14	−1.585	0.795	−1.993	0.184	DPE	28
28	Alpha.Score	DPE	42	14	−1.239	0.650	−1.907	0.218	DPE	28
28	Alpha.Score	DPE	28	21	0.634	1.133	0.560	≥0.25	DPE	28
28	Alpha.Score	DPE	42	21	0.980	1.036	0.946	≥0.25	DPE	28
28	Alpha.Score	DPE	42	28	0.346	0.821	0.421	≥0.25	DPE	28
32	Alpha.Score	DPE	21	14	−0.561	0.412	−1.363	≥0.25	DPE	32
32	Alpha.Score	DPE	28	14	−0.470	0.430	−1.091	≥0.25	DPE	32
32	Alpha.Score	DPE	42	14	−0.357	0.410	−0.871	≥0.25	DPE	32
32	Alpha.Score	DPE	28	21	0.092	0.462	0.199	≥0.25	DPE	32
32	Alpha.Score	DPE	42	21	0.204	0.444	0.460	≥0.25	DPE	32
32	Alpha.Score	DPE	42	28	0.112	0.461	0.243	≥0.25	DPE	32
35	Alpha.Score	DPE	21	14	−0.338	0.242	−1.396	≥0.25	DPE	35
35	Alpha.Score	DPE	28	14	−0.329	0.242	−1.361	≥0.25	DPE	35
35	Alpha.Score	DPE	42	14	−0.233	0.233	−1.000	≥0.25	DPE	35
35	Alpha.Score	DPE	28	21	0.009	0.252	0.035	≥0.25	DPE	35
35	Alpha.Score	DPE	42	21	0.105	0.244	0.430	≥0.25	DPE	35
35	Alpha.Score	DPE	42	28	0.096	0.244	0.394	≥0.25	DPE	35

4D)

Tables

(hide)

Press on tabs to display tables. Scroll to see additional rows.

Capscale

ADONIS2
adonis2(Beta Distance ~ Temperature*DPE); Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	5.218	0.251	14.843	0.001	***
DPE	1.000	0.394	0.019	2.241	0.040	*
Temperature:DPE	2.000	0.548	0.026	1.558	0.110	ns
Residual	83.000	14.589	0.703	NA	NA	NA
Total	88.000	20.748	1.000	NA	NA	NA
canberra
Temperature	2.000	3.573	0.117	5.847	0.001	***
DPE	1.000	0.674	0.022	2.207	0.004	**
Temperature:DPE	2.000	0.913	0.030	1.494	0.015	*
Residual	83.000	25.360	0.831	NA	NA	NA
Total	88.000	30.520	1.000	NA	NA	NA
gunifrac
Temperature	2.000	3.363	0.212	11.847	0.001	***
DPE	1.000	0.261	0.016	1.837	0.102	ns
Temperature:DPE	2.000	0.436	0.028	1.535	0.134	ns
Residual	83.000	11.781	0.744	NA	NA	NA
Total	88.000	15.841	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Exposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temp.DPE	11.000	0.624	0.057	3.261	0.001	**
Residual	77.000	1.339	0.017	NA	NA	NA
canberra
Temp.DPE	11.000	0.316	0.029	2.748	0.005	**
Residual	77.000	0.805	0.010	NA	NA	NA
gunifrac
Temp.DPE	11.000	0.420	0.038	3.232	0.001	**
Residual	77.000	0.909	0.012	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*DPE); Exposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	−0.213	−0.485	0.059	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	−0.258	−0.512	−0.005	0.042	*
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	−0.063	−0.285	0.160	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	−0.059	−0.270	0.152	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.089	−0.319	0.141	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	−0.123	−0.363	0.117	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.142	−0.372	0.088	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.202	−0.418	0.014	0.090	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.170	−0.393	0.052	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	−0.289	−0.511	−0.067	0.002	**
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	−0.192	−0.408	0.024	0.130	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	−0.046	−0.344	0.253	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	0.150	−0.122	0.422	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	0.153	−0.110	0.416	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	0.124	−0.154	0.403	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	0.090	−0.197	0.377	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	0.071	−0.208	0.350	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	0.011	−0.256	0.278	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	0.042	−0.230	0.315	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.077	−0.349	0.196	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	0.021	−0.246	0.288	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	0.196	−0.058	0.449	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	0.199	−0.044	0.443	0.220	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	0.170	−0.091	0.430	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	0.136	−0.133	0.405	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	0.117	−0.144	0.377	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	0.057	−0.191	0.305	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	0.088	−0.165	0.341	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	−0.031	−0.284	0.222	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	0.066	−0.182	0.314	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	0.003	−0.207	0.214	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	−0.026	−0.256	0.204	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	−0.060	−0.300	0.180	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	−0.079	−0.309	0.151	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	−0.139	−0.355	0.077	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	−0.108	−0.330	0.115	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	−0.227	−0.449	−0.004	0.042	*
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	−0.129	−0.345	0.087	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.029	−0.248	0.190	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.063	−0.293	0.166	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.082	−0.301	0.137	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.142	−0.347	0.062	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.111	−0.322	0.100	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.230	−0.441	−0.019	0.021	*
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	−0.133	−0.337	0.072	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	−0.034	−0.281	0.213	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	−0.053	−0.291	0.185	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	−0.113	−0.337	0.111	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	−0.082	−0.312	0.148	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	−0.201	−0.431	0.029	0.148	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	−0.103	−0.327	0.121	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	−0.019	−0.266	0.228	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	−0.079	−0.313	0.155	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	−0.048	−0.288	0.192	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	−0.167	−0.407	0.073	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	−0.069	−0.304	0.165	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	−0.060	−0.284	0.164	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	−0.029	−0.259	0.201	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	−0.148	−0.378	0.082	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	−0.050	−0.274	0.174	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	0.031	−0.185	0.247	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	−0.088	−0.304	0.128	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	0.010	−0.200	0.219	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	−0.119	−0.341	0.103	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	−0.022	−0.238	0.194	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.097	−0.119	0.313	≥0.25	ns
canberra
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	−0.004	−0.215	0.207	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	0.040	−0.156	0.237	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	0.059	−0.113	0.232	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	−0.043	−0.206	0.121	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.044	−0.223	0.134	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	−0.102	−0.288	0.084	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.084	−0.262	0.095	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.097	−0.265	0.070	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.097	−0.269	0.075	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	−0.142	−0.314	0.031	0.215	ns
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	−0.103	−0.271	0.064	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	0.044	−0.187	0.276	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	0.064	−0.148	0.275	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	−0.039	−0.243	0.165	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	−0.040	−0.256	0.176	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	−0.097	−0.320	0.125	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	−0.079	−0.295	0.137	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	−0.093	−0.300	0.114	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	−0.093	−0.304	0.118	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.137	−0.348	0.074	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	−0.099	−0.306	0.108	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	0.019	−0.177	0.216	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	−0.083	−0.272	0.106	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	−0.085	−0.286	0.117	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	−0.142	−0.351	0.067	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	−0.124	−0.326	0.078	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	−0.137	−0.329	0.055	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	−0.137	−0.334	0.059	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	−0.182	−0.378	0.015	0.097	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	−0.144	−0.336	0.049	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	−0.102	−0.266	0.061	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	−0.104	−0.282	0.075	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	−0.161	−0.347	0.025	0.157	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	−0.143	−0.321	0.036	0.247	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	−0.156	−0.324	0.011	0.091	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	−0.156	−0.329	0.016	0.112	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	−0.201	−0.373	−0.028	0.010	**
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	−0.163	−0.330	0.005	0.065	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.001	−0.171	0.168	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.059	−0.237	0.119	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.041	−0.211	0.129	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.054	−0.212	0.104	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.054	−0.218	0.109	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.099	−0.262	0.065	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	−0.060	−0.219	0.098	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	−0.057	−0.249	0.134	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	−0.039	−0.224	0.145	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	−0.053	−0.226	0.121	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	−0.053	−0.231	0.126	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	−0.097	−0.276	0.081	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	−0.059	−0.233	0.115	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	0.018	−0.174	0.210	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	0.005	−0.177	0.186	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	0.005	−0.182	0.191	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	−0.040	−0.226	0.146	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	−0.002	−0.183	0.180	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	−0.013	−0.187	0.160	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	−0.014	−0.192	0.165	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	−0.058	−0.236	0.120	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	−0.020	−0.194	0.154	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	0.000	−0.168	0.167	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	−0.044	−0.212	0.123	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	−0.006	−0.169	0.156	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	−0.044	−0.217	0.128	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	−0.006	−0.174	0.161	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.038	−0.129	0.206	≥0.25	ns
gunifrac
Distance	Temp.DPE	28°C__21DPE	28°C__14DPE	−0.191	−0.415	0.033	0.173	ns
Distance	Temp.DPE	28°C__28DPE	28°C__14DPE	−0.061	−0.270	0.147	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__14DPE	0.018	−0.165	0.201	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__14DPE	−0.024	−0.197	0.150	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__14DPE	−0.027	−0.216	0.163	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__14DPE	−0.100	−0.298	0.098	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__14DPE	−0.069	−0.259	0.120	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__14DPE	−0.131	−0.309	0.047	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__14DPE	−0.137	−0.320	0.046	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__14DPE	−0.198	−0.382	−0.015	0.022	*
Distance	Temp.DPE	35°C__42DPE	28°C__14DPE	−0.134	−0.312	0.044	≥0.25	ns
Distance	Temp.DPE	28°C__28DPE	28°C__21DPE	0.130	−0.116	0.375	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__21DPE	0.209	−0.015	0.433	0.092	ns
Distance	Temp.DPE	32°C__14DPE	28°C__21DPE	0.167	−0.049	0.384	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__21DPE	0.164	−0.065	0.394	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__21DPE	0.091	−0.146	0.327	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__21DPE	0.122	−0.108	0.351	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__21DPE	0.060	−0.161	0.280	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__21DPE	0.054	−0.170	0.278	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__21DPE	−0.007	−0.232	0.217	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__21DPE	0.057	−0.163	0.277	≥0.25	ns
Distance	Temp.DPE	28°C__42DPE	28°C__28DPE	0.079	−0.130	0.288	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__28DPE	0.038	−0.163	0.238	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__28DPE	0.035	−0.180	0.249	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__28DPE	−0.039	−0.260	0.183	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__28DPE	−0.008	−0.222	0.206	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__28DPE	−0.070	−0.274	0.134	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	28°C__28DPE	−0.075	−0.284	0.133	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	28°C__28DPE	−0.137	−0.346	0.072	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	28°C__28DPE	−0.072	−0.277	0.132	≥0.25	ns
Distance	Temp.DPE	32°C__14DPE	28°C__42DPE	−0.041	−0.215	0.132	≥0.25	ns
Distance	Temp.DPE	32°C__21DPE	28°C__42DPE	−0.045	−0.234	0.145	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	28°C__42DPE	−0.118	−0.316	0.080	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	28°C__42DPE	−0.087	−0.277	0.102	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	28°C__42DPE	−0.149	−0.327	0.029	0.190	ns
Distance	Temp.DPE	35°C__21DPE	28°C__42DPE	−0.155	−0.338	0.029	0.182	ns
Distance	Temp.DPE	35°C__28DPE	28°C__42DPE	−0.216	−0.399	−0.033	0.008	**
Distance	Temp.DPE	35°C__42DPE	28°C__42DPE	−0.151	−0.329	0.026	0.173	ns
Distance	Temp.DPE	32°C__21DPE	32°C__14DPE	−0.003	−0.184	0.177	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__14DPE	−0.076	−0.266	0.113	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__14DPE	−0.046	−0.226	0.135	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__14DPE	−0.108	−0.276	0.060	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__14DPE	−0.113	−0.287	0.061	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__14DPE	−0.175	−0.348	−0.001	0.047	*
Distance	Temp.DPE	35°C__42DPE	32°C__14DPE	−0.110	−0.278	0.058	≥0.25	ns
Distance	Temp.DPE	32°C__28DPE	32°C__21DPE	−0.073	−0.277	0.130	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__21DPE	−0.043	−0.238	0.153	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__21DPE	−0.105	−0.289	0.080	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__21DPE	−0.110	−0.300	0.079	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__21DPE	−0.172	−0.361	0.018	0.113	ns
Distance	Temp.DPE	35°C__42DPE	32°C__21DPE	−0.107	−0.292	0.078	≥0.25	ns
Distance	Temp.DPE	32°C__42DPE	32°C__28DPE	0.031	−0.173	0.234	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__28DPE	−0.031	−0.224	0.162	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__28DPE	−0.037	−0.234	0.161	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__28DPE	−0.098	−0.296	0.099	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__28DPE	−0.034	−0.227	0.159	≥0.25	ns
Distance	Temp.DPE	35°C__14DPE	32°C__42DPE	−0.062	−0.247	0.122	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	32°C__42DPE	−0.067	−0.257	0.122	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	32°C__42DPE	−0.129	−0.319	0.060	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	32°C__42DPE	−0.064	−0.249	0.120	≥0.25	ns
Distance	Temp.DPE	35°C__21DPE	35°C__14DPE	−0.005	−0.183	0.173	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__14DPE	−0.067	−0.245	0.111	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__14DPE	−0.002	−0.175	0.170	≥0.25	ns
Distance	Temp.DPE	35°C__28DPE	35°C__21DPE	−0.062	−0.245	0.121	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__21DPE	0.003	−0.175	0.181	≥0.25	ns
Distance	Temp.DPE	35°C__42DPE	35°C__28DPE	0.065	−0.113	0.243	≥0.25	ns

5) Water temperature influences gut microbiome’s sensitivity to parasite exposure

Fig. 5 Water temperature influences gut microbiome response to Pseudocapillaria tomentosa. (A) Simpson’s Index for diversity of parasite unexposed and pre-exposed fish at 0 days post exposure (dpe). Prior to parasite exposure gut microbial alpha-diversity does not significantly differ between fish reared at the same water temperature. (B) Simpson’s Index for diversity of parasite unexposed and exposed fish. Gut microbial alpha-diversity significantly differs between parasite exposed fish reared at 28°C and 32°C water temperature relative to unexposed control fish, but gut microbial alpha-diversity does not differ between parasite unexposed and exposed fish reared at 35°C water temperature. Capscale ordinations based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main and interaction effects of temperature and parasite exposure (treatment) (C) of pre-exposed samples at 0 dpe, and (D) post-exposed samples after 0 dpe. The analysis shows that gut microbiome composition of fish significantly differs between water temperature and parasite exposure in pre- and post-exposed fish. However, the interaction effect between temperature and parasite exposure is not significant in pre-exposed fish, but is significant in post-exposed fish. Ribbons and ellipses indicate 95% confidence interval. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate statistically significant covariates and direction of greatest change in the indicated covariates.

5A)

Tables

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GLM

GLM Results
glm(Alpha.Score ~ Temperature*Treatment); Pre-Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−0.218	0.269	−0.811	≥0.25	ns
Temperature32	0.345	0.379	0.910	≥0.25	ns
Temperature35	0.693	0.384	1.803	0.077	ns
TreatmentExposed	0.274	0.379	0.723	≥0.25	ns
Temperature32:TreatmentExposed	0.311	0.544	0.572	≥0.25	ns
Temperature35:TreatmentExposed	0.041	0.550	0.075	≥0.25	ns
Simpson
(Intercept)	−0.241	0.252	−0.958	≥0.25	ns
Temperature32	0.352	0.355	0.990	≥0.25	ns
Temperature35	0.961	0.367	2.619	0.011	*
TreatmentExposed	0.223	0.355	0.627	≥0.25	ns
Temperature32:TreatmentExposed	0.486	0.513	0.948	≥0.25	ns
Temperature35:TreatmentExposed	0.017	0.525	0.033	≥0.25	ns
Richness
(Intercept)	−0.145	0.270	−0.538	≥0.25	ns
Temperature32	0.550	0.385	1.430	0.158	ns
Temperature35	0.043	0.381	0.112	≥0.25	ns
TreatmentExposed	0.402	0.382	1.050	≥0.25	ns
Temperature32:TreatmentExposed	0.020	0.554	0.036	≥0.25	ns
Temperature35:TreatmentExposed	0.287	0.545	0.526	≥0.25	ns

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Treatment), type = 2); Pre-Exposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	7.264	2.000	0.026	*
Treatment	3.044	1.000	0.081	ns
Temperature:Treatment	0.380	2.000	≥0.25	ns
Simpson
Temperature	14.462	2.000	<0.001	***
Treatment	3.406	1.000	0.065	ns
Temperature:Treatment	1.122	2.000	≥0.25	ns
Richness
Temperature	4.311	2.000	0.116	ns
Treatment	5.067	1.000	0.024	*
Temperature:Treatment	0.340	2.000	≥0.25	ns

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Treatment); Pre-Exposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	Treatment	Exposed	Control	0.274	0.364	0.753	≥0.25	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.585	0.451	1.298	0.194	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.315	0.344	0.917	≥0.25	Treatment	35
Simpson
28	Alpha.Score	Treatment	Exposed	Control	0.223	0.337	0.662	≥0.25	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.709	0.426	1.663	0.096	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.240	0.339	0.708	≥0.25	Treatment	35
Richness
28	Alpha.Score	Treatment	Exposed	Control	0.402	0.361	1.112	≥0.25	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.422	0.411	1.027	≥0.25	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.689	0.400	1.721	0.085	Treatment	35

5B)

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GLM

GLM Results
glm(Alpha.Score ~ Temperature*Treatment); Post-Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−0.441	0.126	−3.499	<0.001	***
Temperature32	−0.064	0.177	−0.364	≥0.25	ns
Temperature35	−0.087	0.182	−0.476	≥0.25	ns
TreatmentExposed	−0.539	0.210	−2.562	0.011	*
Temperature32:TreatmentExposed	0.953	0.280	3.405	<0.001	***
Temperature35:TreatmentExposed	0.561	0.281	1.996	0.047	*
Simpson
(Intercept)	−0.299	0.133	−2.252	0.025	*
Temperature32	−0.059	0.186	−0.319	≥0.25	ns
Temperature35	0.080	0.190	0.419	≥0.25	ns
TreatmentExposed	−0.724	0.225	−3.223	0.001	**
Temperature32:TreatmentExposed	1.106	0.298	3.715	<0.001	***
Temperature35:TreatmentExposed	0.673	0.297	2.270	0.024	*
Richness
(Intercept)	−0.206	0.101	−2.040	0.043	*
Temperature32	−0.048	0.141	−0.343	≥0.25	ns
Temperature35	−0.317	0.147	−2.164	0.032	*
TreatmentExposed	−0.228	0.160	−1.421	0.157	ns
Temperature32:TreatmentExposed	0.829	0.219	3.776	<0.001	***
Temperature35:TreatmentExposed	0.691	0.219	3.155	0.002	**

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Treatment), type = 2); Post-Exposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	5.908	2.000	0.052	ns
Treatment	0.001	1.000	≥0.25	ns
Temperature:Treatment	11.784	2.000	0.003	**
Simpson
Temperature	8.400	2.000	0.015	*
Treatment	0.547	1.000	≥0.25	ns
Temperature:Treatment	14.128	2.000	<0.001	***
Richness
Temperature	11.325	2.000	0.003	**
Treatment	11.824	1.000	<0.001	***
Temperature:Treatment	16.178	2.000	<0.001	***

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Treatment); Post-Exposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	Treatment	Exposed	Control	−0.539	0.280	−1.924	0.054	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.414	0.180	2.298	0.022	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.022	0.111	0.200	≥0.25	Treatment	35
Simpson
28	Alpha.Score	Treatment	Exposed	Control	−0.724	0.290	−2.492	0.013	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.382	0.190	2.013	0.044	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	−0.051	0.130	−0.387	≥0.25	Treatment	35
Richness
28	Alpha.Score	Treatment	Exposed	Control	−0.228	0.223	−1.021	≥0.25	Treatment	28
32	Alpha.Score	Treatment	Exposed	Control	0.601	0.132	4.546	<0.001	Treatment	32
35	Alpha.Score	Treatment	Exposed	Control	0.463	0.092	5.021	<0.001	Treatment	35

5C)

Tables

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Capscale

ADONIS2
adonis2(Beta Distance ~ Temperature*Treatment); Pre-Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	0.871	0.188	7.178	0.001	***
Treatment	1.000	0.244	0.053	4.025	0.010	**
Temperature:Treatment	2.000	0.245	0.053	2.019	0.053	ns
Residual	54.000	3.275	0.707	NA	NA	NA
Total	59.000	4.635	1.000	NA	NA	NA
canberra
Temperature	2.000	1.652	0.132	4.615	0.001	***
Treatment	1.000	0.533	0.043	2.979	0.001	***
Temperature:Treatment	2.000	0.667	0.053	1.864	0.001	***
Residual	54.000	9.665	0.772	NA	NA	NA
Total	59.000	12.517	1.000	NA	NA	NA
gunifrac
Temperature	2.000	0.647	0.153	5.604	0.001	***
Treatment	1.000	0.288	0.068	4.987	0.003	**
Temperature:Treatment	2.000	0.177	0.042	1.537	0.109	ns
Residual	54.000	3.117	0.737	NA	NA	NA
Total	59.000	4.229	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Pre-Exposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temp.Treat	5.000	0.103	0.021	2.738	0.028	*
Residual	54.000	0.405	0.007	NA	NA	NA
canberra
Temp.Treat	5.000	0.014	0.003	0.453	≥0.25	ns
Residual	54.000	0.329	0.006	NA	NA	NA
gunifrac
Temp.Treat	5.000	0.054	0.011	1.564	0.186	ns
Residual	54.000	0.369	0.007	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Treatment); Pre-Exposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.027	−0.088	0.141	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Control	0.095	−0.019	0.210	0.153	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.110	−0.004	0.225	0.065	ns
Distance	Temp.Treat	35°C__Control	28°C__Control	0.015	−0.099	0.130	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.069	−0.045	0.183	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Exposed	0.069	−0.046	0.183	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	0.084	−0.031	0.198	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.011	−0.126	0.103	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	0.042	−0.072	0.157	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.015	−0.100	0.129	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Control	−0.080	−0.195	0.034	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	−0.026	−0.141	0.088	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.095	−0.209	0.019	0.156	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.041	−0.156	0.073	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.054	−0.061	0.168	≥0.25	ns
canberra
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.011	−0.092	0.114	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Control	0.035	−0.068	0.138	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.044	−0.059	0.147	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Control	0.010	−0.094	0.113	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.016	−0.087	0.119	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Exposed	0.024	−0.079	0.127	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	0.033	−0.070	0.136	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.001	−0.104	0.102	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	0.005	−0.098	0.108	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.009	−0.094	0.112	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Control	−0.025	−0.128	0.078	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	−0.019	−0.122	0.084	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.034	−0.137	0.069	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.028	−0.131	0.075	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.006	−0.097	0.109	≥0.25	ns
gunifrac
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.003	−0.106	0.112	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Control	0.051	−0.058	0.160	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.078	−0.032	0.187	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Control	0.000	−0.109	0.110	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.042	−0.067	0.151	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Exposed	0.048	−0.061	0.158	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	0.075	−0.035	0.184	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.002	−0.112	0.107	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	0.039	−0.070	0.148	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.026	−0.083	0.136	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Control	−0.051	−0.160	0.059	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	−0.009	−0.118	0.100	≥0.25	ns
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.077	−0.186	0.032	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.036	−0.145	0.074	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.041	−0.068	0.151	≥0.25	ns

5D)

Tables

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Capscale

ADONIS2
adonis2(Beta Distance ~ Temperature*Treatment); Post-Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	4.123	0.108	17.035	0.001	***
Treatment	1.000	7.411	0.194	61.235	0.001	***
Temperature:Treatment	2.000	3.168	0.083	13.088	0.001	***
Residual	194.000	23.478	0.615	NA	NA	NA
Total	199.000	38.179	1.000	NA	NA	NA
canberra
Temperature	2.000	4.561	0.074	9.502	0.001	***
Treatment	1.000	7.402	0.120	30.840	0.001	***
Temperature:Treatment	2.000	3.014	0.049	6.279	0.001	***
Residual	194.000	46.561	0.757	NA	NA	NA
Total	199.000	61.538	1.000	NA	NA	NA
gunifrac
Temperature	2.000	2.273	0.082	11.945	0.001	***
Treatment	1.000	4.718	0.170	49.577	0.001	***
Temperature:Treatment	2.000	2.273	0.082	11.944	0.001	***
Residual	194.000	18.460	0.666	NA	NA	NA
Total	199.000	27.724	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Post-Exposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temp.Treat	5.000	1.344	0.269	17.882	<0.001	****
Residual	194.000	2.917	0.015	NA	NA	NA
canberra
Temp.Treat	5.000	1.094	0.219	31.758	<0.001	****
Residual	194.000	1.337	0.007	NA	NA	NA
gunifrac
Temp.Treat	5.000	1.225	0.245	29.264	<0.001	****
Residual	194.000	1.624	0.008	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Treatment); Post-Exposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.211	0.119	0.302	<0.001	****
Distance	Temp.Treat	32°C__Control	28°C__Control	−0.015	−0.096	0.066	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.168	0.081	0.255	<0.001	****
Distance	Temp.Treat	35°C__Control	28°C__Control	0.013	−0.070	0.096	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.060	−0.024	0.144	≥0.25	ns
Distance	Temp.Treat	32°C__Control	28°C__Exposed	−0.225	−0.316	−0.135	<0.001	****
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	−0.043	−0.138	0.053	≥0.25	ns
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.197	−0.290	−0.105	<0.001	****
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	−0.150	−0.243	−0.057	<0.001	****
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.183	0.097	0.268	<0.001	****
Distance	Temp.Treat	35°C__Control	32°C__Control	0.028	−0.054	0.110	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	0.075	−0.008	0.158	0.100	ns
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.155	−0.243	−0.067	<0.001	****
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.108	−0.196	−0.019	0.007	**
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.047	−0.038	0.132	≥0.25	ns
canberra
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.217	0.155	0.279	<0.001	****
Distance	Temp.Treat	32°C__Control	28°C__Control	−0.011	−0.066	0.044	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.107	0.048	0.166	<0.001	****
Distance	Temp.Treat	35°C__Control	28°C__Control	0.013	−0.044	0.069	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.073	0.016	0.130	0.004	**
Distance	Temp.Treat	32°C__Control	28°C__Exposed	−0.228	−0.289	−0.167	<0.001	****
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	−0.110	−0.175	−0.045	<0.001	****
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.204	−0.267	−0.142	<0.001	****
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	−0.144	−0.207	−0.081	<0.001	****
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.118	0.060	0.176	<0.001	****
Distance	Temp.Treat	35°C__Control	32°C__Control	0.024	−0.032	0.080	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	0.084	0.028	0.140	<0.001	***
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.094	−0.154	−0.035	<0.001	***
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.034	−0.094	0.026	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.060	0.003	0.118	0.035	*
gunifrac
Distance	Temp.Treat	28°C__Exposed	28°C__Control	0.197	0.129	0.266	<0.001	****
Distance	Temp.Treat	32°C__Control	28°C__Control	−0.023	−0.083	0.037	≥0.25	ns
Distance	Temp.Treat	32°C__Exposed	28°C__Control	0.141	0.077	0.206	<0.001	****
Distance	Temp.Treat	35°C__Control	28°C__Control	−0.011	−0.073	0.051	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	28°C__Control	0.054	−0.009	0.117	0.134	ns
Distance	Temp.Treat	32°C__Control	28°C__Exposed	−0.220	−0.288	−0.153	<0.001	****
Distance	Temp.Treat	32°C__Exposed	28°C__Exposed	−0.056	−0.128	0.015	0.211	ns
Distance	Temp.Treat	35°C__Control	28°C__Exposed	−0.209	−0.278	−0.140	<0.001	****
Distance	Temp.Treat	35°C__Exposed	28°C__Exposed	−0.143	−0.213	−0.074	<0.001	****
Distance	Temp.Treat	32°C__Exposed	32°C__Control	0.164	0.100	0.228	<0.001	****
Distance	Temp.Treat	35°C__Control	32°C__Control	0.012	−0.050	0.073	≥0.25	ns
Distance	Temp.Treat	35°C__Exposed	32°C__Control	0.077	0.015	0.139	0.006	**
Distance	Temp.Treat	35°C__Control	32°C__Exposed	−0.152	−0.218	−0.087	<0.001	****
Distance	Temp.Treat	35°C__Exposed	32°C__Exposed	−0.087	−0.153	−0.021	0.003	**
Distance	Temp.Treat	35°C__Exposed	35°C__Control	0.065	0.002	0.129	0.039	*

6) Water temperature moderates the impact of infection burden on fish gut microbiomes

Fig. 6 Exposure to Pseudocapillaria tomentosa differentially impacts gut microbiome depending on water temperature. (A) Simpson’s Index for diversity of parasite exposed fish. Gut microbial alpha-diversity does not significantly differ between fish reared at the same water temperature with presence or absence of infection. (B) Capscale ordination based on the Canberra dissimilarity of gut microbiome composition of parasite exposed fish. The analysis shows that gut microbiome composition significantly differs between positively infected fish reared at different water temperatures. (C) Total worm count is positively correlated with lowest or highest alpha diversity scores in fish with detectable parasites. Points are colored by temperature and filled by cluster (High = purple, Low = orange, Other = white), and samples with a worm count equal to 0 are transparent. (D) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition of water temperature and worm counts. Points are colored by temperature and filled by cluster (High = purple, Low = orange, Other = white), and samples with a worm count equal to 0 are transparent. The analysis shows that gut microbiome composition significantly differs between water temperature and total worm count, and samples labeled as “High” or “Low” form distinct clusters. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate statistically significant covariates and direction of greatest change in the indicated covariates.

6A)

Tables

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GLM

GLM Results
glm(Alpha.Score ~ Temperature*Pathology.Results); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	−1.090	0.429	−2.544	0.013	*
Temperature32	0.889	0.508	1.749	0.084	ns
Temperature35	0.693	0.468	1.482	0.142	ns
Pathology.Resultspositive	0.223	0.512	0.436	≥0.25	ns
Temperature32:Pathology.Resultspositive	0.125	0.641	0.195	≥0.25	ns
Temperature35:Pathology.Resultspositive	−1.122	1.392	−0.806	≥0.25	ns
Simpson
(Intercept)	−1.265	0.458	−2.760	0.007	**
Temperature32	1.105	0.536	2.061	0.042	*
Temperature35	1.055	0.495	2.130	0.036	*
Pathology.Resultspositive	0.375	0.541	0.694	≥0.25	ns
Temperature32:Pathology.Resultspositive	0.054	0.669	0.081	≥0.25	ns
Temperature35:Pathology.Resultspositive	−1.154	1.337	−0.863	≥0.25	ns
Richness
(Intercept)	−0.421	0.329	−1.279	0.204	ns
Temperature32	0.483	0.405	1.194	0.236	ns
Temperature35	−0.002	0.367	−0.005	≥0.25	ns
Pathology.Resultspositive	−0.390	0.407	−0.957	≥0.25	ns
Temperature32:Pathology.Resultspositive	0.310	0.526	0.589	≥0.25	ns
Temperature35:Pathology.Resultspositive	−0.221	1.125	−0.197	≥0.25	ns

ANOVA

ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Exposed fish
term	statistic	df	p.value	sig
Shannon
Temperature	9.895	2.000	0.007	**
Pathology.Results	0.569	1.000	≥0.25	ns
Temperature:Pathology.Results	0.997	2.000	≥0.25	ns
Simpson
Temperature	13.663	2.000	0.001	**
Pathology.Results	1.134	1.000	≥0.25	ns
Temperature:Pathology.Results	0.999	2.000	≥0.25	ns
Richness
Temperature	8.625	2.000	0.013	*
Pathology.Results	0.840	1.000	≥0.25	ns
Temperature:Pathology.Results	0.495	2.000	≥0.25	ns

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Exposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	Pathology.Results	positive	negative	0.223	0.732	0.305	≥0.25	Pathology.Results	28
32	Alpha.Score	Pathology.Results	positive	negative	0.348	0.383	0.908	≥0.25	Pathology.Results	32
35	Alpha.Score	Pathology.Results	positive	negative	−0.899	0.659	−1.365	0.172	Pathology.Results	35
Simpson
28	Alpha.Score	Pathology.Results	positive	negative	0.375	0.754	0.497	≥0.25	Pathology.Results	28
32	Alpha.Score	Pathology.Results	positive	negative	0.429	0.394	1.089	≥0.25	Pathology.Results	32
35	Alpha.Score	Pathology.Results	positive	negative	−0.779	0.692	−1.125	≥0.25	Pathology.Results	35
Richness
28	Alpha.Score	Pathology.Results	positive	negative	−0.390	0.608	−0.641	≥0.25	Pathology.Results	28
32	Alpha.Score	Pathology.Results	positive	negative	−0.080	0.301	−0.267	≥0.25	Pathology.Results	32
35	Alpha.Score	Pathology.Results	positive	negative	−0.611	0.552	−1.107	≥0.25	Pathology.Results	35

6B)

Tables

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Capscale

ADONIS2
adonis2(Beta Distance ~ Temperature*Pathology.Results); Exposed fish
term	df	SumOfSqs	R2	statistic	p.value	sig
bray
Temperature	2.000	5.218	0.251	14.569	0.001	***
Pathology.Results	1.000	0.263	0.013	1.471	0.185	ns
Temperature:Pathology.Results	2.000	0.404	0.019	1.127	≥0.25	ns
Residual	83.000	14.863	0.716	NA	NA	NA
Total	88.000	20.748	1.000	NA	NA	NA
canberra
Temperature	2.000	3.573	0.117	5.751	0.001	***
Pathology.Results	1.000	0.488	0.016	1.570	0.031	*
Temperature:Pathology.Results	2.000	0.679	0.022	1.093	≥0.25	ns
Residual	83.000	25.781	0.845	NA	NA	NA
Total	88.000	30.520	1.000	NA	NA	NA
gunifrac
Temperature	2.000	3.363	0.212	11.651	0.001	***
Pathology.Results	1.000	0.234	0.015	1.622	0.149	ns
Temperature:Pathology.Results	2.000	0.264	0.017	0.915	≥0.25	ns
Residual	83.000	11.980	0.756	NA	NA	NA
Total	88.000	15.841	1.000	NA	NA	NA

Dispersion (ANOVA)

ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature*Pathology.Results); Exposed fish
term	df	sumsq	meansq	statistic	p.value	sig
bray
Temp.Path	5.000	0.520	0.104	4.652	<0.001	***
Residual	83.000	1.855	0.022	NA	NA	NA
canberra
Temp.Path	5.000	0.625	0.125	13.830	<0.001	****
Residual	83.000	0.750	0.009	NA	NA	NA
gunifrac
Temp.Path	5.000	0.436	0.087	6.954	<0.001	****
Residual	83.000	1.040	0.013	NA	NA	NA

Dispersion (Tukey)

Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Pathology.Results); Exposed fish
.y.	term	group1	group2	estimate	conf.low	conf.high	adj.p.value	sig
bray
Distance	Temp.Path	28°C__pos	28°C__neg	0.022	−0.165	0.209	≥0.25	ns
Distance	Temp.Path	32°C__neg	28°C__neg	−0.086	−0.277	0.105	≥0.25	ns
Distance	Temp.Path	32°C__pos	28°C__neg	0.015	−0.176	0.206	≥0.25	ns
Distance	Temp.Path	35°C__neg	28°C__neg	−0.126	−0.298	0.046	≥0.25	ns
Distance	Temp.Path	35°C__pos	28°C__neg	−0.434	−0.896	0.029	0.079	ns
Distance	Temp.Path	32°C__neg	28°C__pos	−0.108	−0.263	0.046	≥0.25	ns
Distance	Temp.Path	32°C__pos	28°C__pos	−0.007	−0.161	0.148	≥0.25	ns
Distance	Temp.Path	35°C__neg	28°C__pos	−0.148	−0.278	−0.017	0.017	*
Distance	Temp.Path	35°C__pos	28°C__pos	−0.456	−0.905	−0.007	0.044	*
Distance	Temp.Path	32°C__pos	32°C__neg	0.101	−0.058	0.261	≥0.25	ns
Distance	Temp.Path	35°C__neg	32°C__neg	−0.039	−0.175	0.096	≥0.25	ns
Distance	Temp.Path	35°C__pos	32°C__neg	−0.348	−0.798	0.103	0.226	ns
Distance	Temp.Path	35°C__neg	32°C__pos	−0.141	−0.277	−0.005	0.038	*
Distance	Temp.Path	35°C__pos	32°C__pos	−0.449	−0.899	0.001	0.051	ns
Distance	Temp.Path	35°C__pos	35°C__neg	−0.308	−0.751	0.135	≥0.25	ns
canberra
Distance	Temp.Path	28°C__pos	28°C__neg	0.045	−0.074	0.164	≥0.25	ns
Distance	Temp.Path	32°C__neg	28°C__neg	−0.123	−0.244	−0.002	0.045	*
Distance	Temp.Path	32°C__pos	28°C__neg	−0.042	−0.164	0.079	≥0.25	ns
Distance	Temp.Path	35°C__neg	28°C__neg	−0.105	−0.214	0.005	0.068	ns
Distance	Temp.Path	35°C__pos	28°C__neg	−0.588	−0.883	−0.294	<0.001	****
Distance	Temp.Path	32°C__neg	28°C__pos	−0.168	−0.267	−0.070	<0.001	****
Distance	Temp.Path	32°C__pos	28°C__pos	−0.088	−0.186	0.010	0.107	ns
Distance	Temp.Path	35°C__neg	28°C__pos	−0.150	−0.233	−0.067	<0.001	****
Distance	Temp.Path	35°C__pos	28°C__pos	−0.634	−0.919	−0.348	<0.001	****
Distance	Temp.Path	32°C__pos	32°C__neg	0.080	−0.021	0.182	0.199	ns
Distance	Temp.Path	35°C__neg	32°C__neg	0.018	−0.068	0.105	≥0.25	ns
Distance	Temp.Path	35°C__pos	32°C__neg	−0.465	−0.752	−0.179	<0.001	***
Distance	Temp.Path	35°C__neg	32°C__pos	−0.062	−0.149	0.024	≥0.25	ns
Distance	Temp.Path	35°C__pos	32°C__pos	−0.546	−0.832	−0.259	<0.001	****
Distance	Temp.Path	35°C__pos	35°C__neg	−0.484	−0.765	−0.202	<0.001	****
gunifrac
Distance	Temp.Path	28°C__pos	28°C__neg	0.011	−0.129	0.151	≥0.25	ns
Distance	Temp.Path	32°C__neg	28°C__neg	−0.091	−0.234	0.052	≥0.25	ns
Distance	Temp.Path	32°C__pos	28°C__neg	−0.014	−0.157	0.129	≥0.25	ns
Distance	Temp.Path	35°C__neg	28°C__neg	−0.129	−0.258	0.000	0.049	*
Distance	Temp.Path	35°C__pos	28°C__neg	−0.411	−0.758	−0.065	0.011	*
Distance	Temp.Path	32°C__neg	28°C__pos	−0.102	−0.218	0.014	0.115	ns
Distance	Temp.Path	32°C__pos	28°C__pos	−0.025	−0.141	0.091	≥0.25	ns
Distance	Temp.Path	35°C__neg	28°C__pos	−0.140	−0.238	−0.043	<0.001	***
Distance	Temp.Path	35°C__pos	28°C__pos	−0.423	−0.759	−0.087	0.006	**
Distance	Temp.Path	32°C__pos	32°C__neg	0.077	−0.042	0.196	≥0.25	ns
Distance	Temp.Path	35°C__neg	32°C__neg	−0.038	−0.140	0.064	≥0.25	ns
Distance	Temp.Path	35°C__pos	32°C__neg	−0.321	−0.658	0.017	0.072	ns
Distance	Temp.Path	35°C__neg	32°C__pos	−0.115	−0.217	−0.013	0.017	*
Distance	Temp.Path	35°C__pos	32°C__pos	−0.398	−0.735	−0.060	0.011	*
Distance	Temp.Path	35°C__pos	35°C__neg	−0.283	−0.614	0.049	0.140	ns

6C)

Tables

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GLM

GLM Results
glm(Total.Worm.Count ~ Alpha.Score*Cluster); Exposed fish
term	estimate	std.error	statistic	p.value	p.adj.sig
Shannon
(Intercept)	0.000	0.778	0.000	≥0.25	ns
Alpha.Score	0.000	1.808	0.000	≥0.25	ns
ClusterLow	6.524	1.067	6.115	<0.001	****
ClusterHigh	−2.610	3.512	−0.743	≥0.25	ns
Alpha.Score:ClusterLow	−10.090	4.999	−2.018	0.047	*
Alpha.Score:ClusterHigh	13.706	4.766	2.876	0.005	**
Simpson
(Intercept)	0.000	0.741	0.000	≥0.25	ns
Alpha.Score	0.000	1.605	0.000	≥0.25	ns
ClusterLow	6.842	1.041	6.573	<0.001	****
ClusterHigh	−3.247	3.900	−0.833	≥0.25	ns
Alpha.Score:ClusterLow	−12.165	4.647	−2.618	0.011	*
Alpha.Score:ClusterHigh	14.198	5.090	2.789	0.007	**
Richness
(Intercept)	0.000	0.914	0.000	≥0.25	ns
Alpha.Score	0.000	1.983	0.000	≥0.25	ns
ClusterLow	6.105	1.294	4.717	<0.001	****
ClusterHigh	7.362	2.644	2.784	0.007	**
Alpha.Score:ClusterLow	−3.491	4.463	−0.782	≥0.25	ns
Alpha.Score:ClusterHigh	0.772	4.163	0.185	≥0.25	ns

ANOVA

ANOVA of GLM
ANOVA(GLM(Total.Worm.Count ~ Alpha.Score*Cluster), type = 2); Exposed fish
term	statistic	df	p.value	sig
Shannon
Alpha.Score	0.143	1.000	≥0.25	ns
Cluster	162.212	2.000	<0.001	****
Alpha.Score:Cluster	14.204	2.000	<0.001	***
Simpson
Alpha.Score	0.002	1.000	≥0.25	ns
Cluster	168.513	2.000	<0.001	****
Alpha.Score:Cluster	16.419	2.000	<0.001	***
Richness
Alpha.Score	0.066	1.000	≥0.25	ns
Cluster	145.539	2.000	<0.001	****
Alpha.Score:Cluster	0.741	2.000	≥0.25	ns

Tukey

Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Total.Worm.Count ~ Alpha.Score*Cluster); Exposed fish
Temperature	.y.	term	group1	group2	estimate	std.error	statistic	adj.p.value	Variable	Group
Shannon
28	Alpha.Score	Cluster	Low	Other	−1.594	0.642	−2.483	0.034	Cluster	28
28	Alpha.Score	Cluster	High	Other	2.844	0.672	4.233	<0.001	Cluster	28
28	Alpha.Score	Cluster	High	Low	4.438	0.773	5.741	<0.001	Cluster	28
32	Alpha.Score	Cluster	Low	Other	−1.391	0.495	−2.808	0.013	Cluster	32
32	Alpha.Score	Cluster	High	Other	1.147	0.333	3.443	0.002	Cluster	32
32	Alpha.Score	Cluster	High	Low	2.538	0.528	4.811	<0.001	Cluster	32
35	Alpha.Score	Cluster	Low	Other	−0.899	0.659	−1.365	0.172	Cluster	35
Simpson
28	Alpha.Score	Cluster	Low	Other	−1.487	0.656	−2.266	0.060	Cluster	28
28	Alpha.Score	Cluster	High	Other	2.970	0.665	4.466	<0.001	Cluster	28
28	Alpha.Score	Cluster	High	Low	4.457	0.767	5.810	<0.001	Cluster	28
32	Alpha.Score	Cluster	Low	Other	−1.233	0.487	−2.531	0.029	Cluster	32
32	Alpha.Score	Cluster	High	Other	1.264	0.354	3.573	<0.001	Cluster	32
32	Alpha.Score	Cluster	High	Low	2.497	0.528	4.729	<0.001	Cluster	32
35	Alpha.Score	Cluster	Low	Other	−0.779	0.692	−1.125	≥0.25	Cluster	35
Richness
28	Alpha.Score	Cluster	Low	Other	−1.430	0.589	−2.429	0.040	Cluster	28
28	Alpha.Score	Cluster	High	Other	1.367	0.653	2.095	0.090	Cluster	28
28	Alpha.Score	Cluster	High	Low	2.797	0.692	4.043	<0.001	Cluster	28
32	Alpha.Score	Cluster	Low	Other	−1.183	0.387	−3.052	0.006	Cluster	32
32	Alpha.Score	Cluster	High	Other	0.429	0.278	1.543	≥0.25	Cluster	32
32	Alpha.Score	Cluster	High	Low	1.611	0.410	3.934	<0.001	Cluster	32
35	Alpha.Score	Cluster	Low	Other	−0.611	0.552	−1.107	≥0.25	Cluster	35

6D)

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Capscale

Dispersion (ANOVA)

Dispersion (Tukey)

7) Gut microbial abundance is significantly associated with environmental conditions and stressors

7A)

Fig. 7 A heatmap of model coefficient values of the top 50 statistically significant abundant gut microbial taxa identified by MaAsLin2. The color of each cell represents the coefficient value and direction (red is positive, blue is negative). A “+” or “-” indicates a statistically significant association was observed between taxon abundance and a covariate. Gray colored cells indicate a significant effect was not observed.

Taxon	Temp: 32°C	Temp: 35°C	Time (DPE)	Cluster: Low	Parasite exposed	Infection present	Infection burden
Actinobacteriota
IMCC26207	+		-	-	-
Nocardiaceae Genus	+	+	+
PeM15 Genus	+		-	-	-
Bacteroidota
Barnesiellaceae Genus			-		-
Cloacibacterium		+	-
Flavobacterium	-	-	-	-	+
Fluviicola		-	-		-	-
Microscillaceae Genus	+	+	+	-
Terrimonas				-		+	+
env OPS 17 Genus		-		-	-
Chloroflexi
JG30 KF CM45 Genus			+	-	+	+
Cyanobacteria
Candidatus Obscuribacter	-	-	+	-	+	+
Obscuribacteraceae Genus	+		+	-		+	-
Vampirovibrionaceae Genus			+	-	+	+
Desulfobacterota
Desulfobacterota Genus				-	+	+
Firmicutes
Clostridium sensu stricto 3			+	-	+	+
Gemmatimonadota
Gemmatimonadaceae Genus		-		-	+	+
Myxococcota
Sandaracinaceae Genus	+	+	+	-	+	+
NB1-j
NB1 j Genus		+	+	-	+	+
Nitrospirota
Nitrospira		-	+	-	+	+
Patescibacteria
Saccharimonadales Genus	+	+	+	-	+
Planctomycetota
CL500 3			+	-	-	+
Gemmata		+	+	-	+	+
Pirellulaceae Genus	+	-	+	-	-
Planctomycetales Genus			+	-	+	+
Planctomycetes Genus			+	-	+	+
Rhodopirellula				-	+	+	-
SM1A02			+	-	+	+
Telmatocola			+	-	+	+
Proteobacteria
A0839 Genus	+			-	+	+
Alphaproteobacteria Genus		+	+	-	+	+
Bauldia	+		+	-	+	+
Beijerinckiaceae Genus		-	+	-		+	-
Bosea	-	-	-	-
Candidatus Alysiosphaera	+		+	-	+	+
Chitinibacter		+	+		+		-
Comamonadaceae Genus			-	-	+	+
Gammaproteobacteria Genus	-	-	+	-		+
Luteimonas	-			-		+	+
Methyloligellaceae Genus	+	+	+	-	+	+
Nordella		-		-	+	+
Plesiomonas	+	+			-	-	-
Pseudomonadales Genus	-	-	-	-	-	-
Rhizobacter				-	+	+	+
Rhizobiales Incertae Sedis Genus				-		+
Rhodobacteraceae Genus				-	+	+
Rhodovastum		-		-
Steroidobacteraceae Genus			+	-	+	+
Xanthobacteraceae Genus	-	-	+	-		+
Verrucomicrobiota
Oikopleura			+	-	+	+